CN114796105B

CN114796105B - Stable AST-3424 injection preparation and preparation method thereof

Info

Publication number: CN114796105B
Application number: CN202210383334.6A
Authority: CN
Inventors: 段建新
Original assignee: Shenzhen Ascentawits Pharmaceutical Technology Co ltd
Current assignee: Shenzhen Ascentawits Pharmaceutical Technology Co ltd
Priority date: 2019-07-15
Filing date: 2020-07-14
Publication date: 2023-09-01
Anticipated expiration: 2040-07-14
Also published as: JP7320086B2; CN112220742A; TW202116298A; BR112021026483A2; JP2022541121A; CN112469394A; EP4000601A4; KR20220015456A; EP4000601A1; CA3147124A1; AU2020314494A1; WO2021008520A1; CN112469394B; AU2020314494A8; IL289652A; US20230181606A1; TWI844703B; CN114796105A

Abstract

A stable AST-3424 injection preparation is a solution containing 0.1-200mg/ml or 1-200mg/ml AST-3424 raw material medicine, wherein the solvent of the solution contains monohydric alcohol of C2-C8. A method of preparation comprising the operations of: dissolving AST-3424 raw material medicine and partial ethanol with a prescription amount for the first time; adding the propylene glycol with the prescription amount for dissolving and preparing for the second time; adding the rest ethanol with the prescription amount, mixing and dissolving to obtain solution containing 1-200mg/ml AST-3424 crude drug. The injection for AST-3424 has water, glucose solution, ethanol and propylene glycol solute, and consists of AST-3424 material medicine, isotonic regulating reagent and pH regulator, and has concentration of AST-3424 material medicine of 0.001-1.000mg/ml and pH of 6.8-10.5 and isotonic solution.

Description

Stable AST-3424 injection preparation and preparation method thereof

The application relates to a Chinese patent application with the application number of 202080001484.5 and the application name of stable AST-3424 injection preparation and the preparation method, which are filed in 7 months and 14 days in 2020.

Technical Field

The invention relates to the research and development of injection of a compound disclosed in patent application PCT/US2016/021581, publication number WO2016145092A1, corresponding to Chinese application number 2016800150788 and publication number CN107530556A, and belongs to the field of the research and development of preparations of cancer treatment compounds.

Background

DNA alkylating cancer therapeutic drug AST-3424 (see patent application: DNA alkylating agent, corresponding to PCT application No. PCT/US2016/021581, publication No. WO2016/145092, corresponding to China application No. 2016800150788, publication No. CN107530556A, disclosed compound TH2870, (R) -1- (3- (3-N, N-dimethylaminocarbonyl) phenoxy-4-nitrophenyl) -1-ethyl-N, N '-bis (ethylenephosphoramidate), composition and method of use and preparation thereof, corresponding to PCT application No. PCT/US2016/062114, publication No. WO2017087428A1, corresponding to PCT application No. 2016800446081, publication No. CN108290911A, chinese name (S) -1- (3- (3-N, N-dimethylaminocarbonyl) phenoxy-4-nitrophenyl) -1-ethyl-N, N' -bis (ethylenephosphoramidate, also referred to as I-24, OB-70) and structure of CAS No. 2097713-2825A), which was developed by I company, is characterized by the following structure:

Industry authoritative documents (KathrynEvans, jianXinDuan, taraPritchard, etal.OBI-3424,anove lAKR1C3-activatedpro drug, exhibitdescript macleaya Smith, abstract LB-B16: the AKR1C 3-acteve Provantagon ProugI-3424 ExertsProfondInViviceEffcient BoschafricailInflutsprailInflutsatpres ModelModelSOftT-CellAcutmeutmeomphaluja; aPedia, preclacinica Testingcondustium Studiy [ C ], AACR-NCI-EORTCInterationalConference: molecular TargetsandC and therapeutic applications; october26-30,2017; philadelphia, pa., DOI: 10.1158/1535-7163.) confirmed that the compound was a broad-spectrum small-molecule anticancer prodrug, having therapeutic effects on a variety of solid tumors and hematological tumors.

For subsequent clinical trials, it is necessary to prepare the appropriate dosage form for human administration: usually administered orally or by injection.

The substance is found to be light yellow liquid in the synthetic preparation process, and various difficulties exist in the aspects of storage, transportation and preparation: because of the amide and phosphate structures, the development of the tablet and oral liquid forms of the oral administration forms is inconvenient, however, the preliminary experiments of research and development teams find that the conventional injection taking water as a solvent is insufficient in stability, and cannot meet the requirements of the subsequent multi-center and multi-sample long-term clinical experiments and commercial production and sales.

Disclosure of Invention

The application provides a stable AST-3424 injection preparation, a preparation method and related technologies, and the injection preparation provided by the technology meets the requirements of long-term clinical tests and commercial production and sales through tests. Specifically, the application discloses the following technical scheme.

The application discloses an AST-3424 injection preparation in a storage stable concentrated form, which is a concentrated injection of a pharmaceutical composition, wherein the composition needs to be diluted before administration and needs to be diluted or prepared by medical staff or pharmacist or pharmacy or factory when in use according to requirements.

The concentrated injection of the present application may be diluted with any pharmaceutically acceptable solution of isotonic adjusting agent (as a solute) and water for injection (as a solvent).

Suitable isotonic agents include, but are not limited to, sodium chloride, dextrose, sucrose, fructose, xylitol, glycerol, sorbitol, mannitol, potassium chloride, mannose, calcium chloride, magnesium chloride and other inorganic salts in anhydrous or aqueous form. Preferably, the osmolyte is glucose or sodium chloride.

A stable AST-3424 injection preparation is a solution containing 0.1-200mg/ml AST-3424 bulk drug, wherein the solvent of the solution contains monohydric alcohol of C2-C8.

Further, the solvent of the solution is a liquid solvent obtained by mixing monohydric alcohol containing C2-C8 and dihydric alcohol or trihydric alcohol containing C2-C8.

Obviously, to further enhance stability against time and heat, other substances may be added which are not harmful and which do not react with the AST-3424 drug substance, generally these substances include:

the protective gas is used for greatly reducing the content of gases with certain activity such as O2, CO2 and the like in the injection after vacuumizing the injection, and then filling inert gases which do not react with the bulk drugs, monohydric alcohol, dihydric alcohol or trihydric alcohol, such as N2 or other inert gases, wherein the protective gases can be dissolved in the injection;

antioxidants, the stability is further improved by adding an appropriate amount of antioxidants (such as vitamin E, vitamin C or glutathione) to the injection.

To improve compliance with patient use, a pharmacopoeia or prescription set of drugs with anesthetic or analgesic effect may be added correspondingly.

The injection disclosed by the invention can also contain another therapeutic agent to form a compound preparation, and plays a role in enhancing the therapeutic effect after synergism. In particular, these drugs are recommended as drugs that modulate the expression levels of AKR1C3 enzymes or the corresponding genes.

In some cases, in order to cope with special environments, the injection disclosed by the invention can be added with antibacterial agents and antifungal agents.

In addition, an electrolyte such as salt NaCl or KCl, an aqueous salt for adjusting the balance of electrolyte and acid and alkali, and an osmotic pressure, which are often used in an injection, glucose, amino acid, or the like may be added as appropriate.

Substances for adjusting pH include alkali metal salts and alkaline earth metal salts of weak acids such as carbonic acid, phosphoric acid, citric acid and acetic acid, and inorganic bases such as alkali metal hydroxides and alkaline earth metal hydroxides such as Na and K: sodium citrate, potassium citrate, sodium acetate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate. Some embodiments of the present disclosure demonstrate that certain organic amine organic bases may cause instability of the injection, and although the mechanism of instability is not yet clear, such test results instruct or inspire that it is not desirable to add organic amine organic bases, such as triethylamine, triethanolamine, and other organic amines, to the results of non-rigorous stability tests.

Obviously, the addition amount of the protective gas, the antioxidant, the drug with anesthesia or pain relieving effect, the antibacterial agent, the antifungal agent and the like is very small, and the additive does not influence the solubility and other properties of the AST-3424 bulk drug or can enhance the stability; the other therapeutic agent, substances for regulating water salt, electrolyte and acid-base balance, osmotic pressure and substances for regulating pH are added according to the purposes, prescription amount, pharmacopoeia, prescription set and other practical conditions.

Monohydric C2-C8 alcohols, including straight chain, branched, cyclic types of fatty alcohols, aromatic alcohols (excluding phenols and higher activity benzyl alcohol).

C2-C8 diols or triols, including straight chain, branched, cyclic types of fatty alcohols, aromatic alcohols (excluding phenols and higher activity benzyl alcohols).

It is obvious that the mixed solvent of the monohydric alcohol of C2-C8 and the dihydric alcohol or the trihydric alcohol of C2-C8 defined in the invention means that the monohydric alcohol and the dihydric alcohol or the trihydric alcohol are respectively liquid at normal temperature and pressure or the mixed solvent is liquid after being mixed. The polyol is preferably ethylene glycol, propylene glycol, glycerol, mannitol, sorbitol or mixtures thereof.

Further, the solvent of the solution is a liquid or semi-liquid mixture of monohydric alcohol containing C2-C8 and water-soluble high polymer suitable for medical use.

In pharmaceutical formulations, liquid or semi-liquid mixtures of pharmaceutically suitable water-soluble high molecular polymers include a wide variety of: polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, poloxamer, polysorbate, dextran, and the like.

The monohydric alcohol (preferably ethanol) contained in the injection preparation provided by the invention has the main effect of dissolving and diluting viscous oily AST-3424 bulk drug (the monohydric alcohol such as ethanol has good solubility to the bulk drug in the synthesis process, but experiments prove that the stability of the ethanol solution is relatively poor).

Because of the poor stability of the single monohydric alcohol solution such as ethanol, the concentration in the solution is presumably changed due to volatility; or because the ethanol molecules of the solvent are too active, the molecular movement of the bulk drug is not hindered, and the change is faster, so that the stability is poor.

After dihydric alcohol or trihydric alcohol, water-soluble high molecular polymer and the like are added into monohydric alcohol, the solution becomes viscous, the thickening effect is achieved, and meanwhile, the added substances can also block the movement of the raw material medicine molecules, so that the stability is enhanced.

Further, the solvent of the stable AST-3424 injection is a mixture of monohydric alcohol of C2-C4 and dihydric alcohol of C2-C3, more preferably a mixture of ethanol and propylene glycol.

Further, the volume ratio of the monohydric alcohol in the mixed solvent of the stable AST-3424 injection is not less than 50%.

Preferably, the mixed solvent of the stable AST-3424 injection consists of 75% of ethanol and 25% of propylene glycol by volume ratio.

Considering various factors such as stability, convenience in use and the like, the stable AST-3424 injection disclosed by the invention is a solution containing 10mg/ml of AST-3424 bulk drug, the concentration is a recommended value of a research and development team, and the stable AST-3424 injection shows better properties in comparison of experiments.

Further, the stable AST-3424 injection disclosed in the document is free of water, and the water content is controlled within 0.5% by mass.

Obviously, no water is added in the preparation process, and other reagents, solvents and the like used are anhydrous grade reagents, such as absolute ethyl alcohol and absolute propylene glycol, namely the water content needs to be controlled in the preparation process of the injection, in theory, the lower the water content of the injection is, the better the stability is, but the research and development team considers the technical difficulty and the realization of industrial mass production, and the water content (measured by a Karl Fischer method) is better within 0.5 percent of the mass ratio.

A stable AST-3424 injection consists of 0.75ml of absolute ethyl alcohol, 0.25ml of absolute propylene glycol and 10mg of AST-3424 bulk drug.

The 1.0ml injection containing 10mgAST-3424 is a specific example, which is poured into a medium borosilicate glass tube (brown) injection bottle with 2ml (or 5 ml) volume, then filled with inert protective gas, covered with rubber plug and sealed with aluminum plastic combined cover for antibiotic bottle.

Obviously, the above 0.75ml of anhydrous ethanol and 0.25ml of anhydrous propylene glycol change in volume (have been demonstrated to decrease) after mixing, so reasonable volume changes are also well known to those skilled in the art (researchers or medical staff in the fields of medicine development, organic chemistry synthesis, formulation development, etc.).

In particular, the concentration of AST-3424 raw material medicine of the injection in this embodiment is calibrated to be 10mg/ml, and similarly, the content actually tested in the medical standard also changes, and the concentration is considered to be qualified in the corresponding range of the pharmacopoeia, the prescription set or the medicine standard, that is, the concentration in the corresponding range is technically equivalent to the calibrated concentration of 10 mg/ml.

The stable AST-3424 injection preparation product comprises a packaging container and an AST-3424 injection preparation contained in the container, wherein the injection preparation is prepared from 1-200mg, preferably 10mg and 20mg of AST-3424 raw material medicine in the product, and the injection preparation is filled in a light-resistant glass bottle made of medium borosilicate glass filled with protective gas.

A stable AST-3424 injection preparation product consists essentially of 0.75ml of absolute ethyl alcohol, 0.25ml of absolute propylene glycol and 10mg of AST-3424 bulk drug, and is filled in a light-resistant glass bottle made of medium borosilicate glass with volumes of 2, 5 and 10ml filled with protective gas.

A method for preparing stable AST-3424 injection, which comprises the following operations:

dissolving AST-3424 raw material medicine and partial ethanol with a prescription amount for the first time;

Adding the propylene glycol with the prescription amount for dissolving and preparing for the second time;

adding the rest ethanol with the prescription amount, mixing and dissolving to obtain solution containing 1-200mg/ml or 1-200mg/ml AST-3424 crude drug.

In the preparation method, AST-3424 raw material medicines and part of ethanol with the prescription amount are dissolved for the first time to prepare a relatively concentrated solution, and then propylene glycol and the rest ethanol with the prescription amount are added for the second time to dilute.

The procedure is based on the nature of the viscous oil of the AST-3424 drug substance. Preferably, the ethanol formulated for the first dissolution is 50% of the prescribed amount.

Or dissolving AST-3424 with ethanol for the first time; adding propylene glycol with the prescribed amount for the second dissolution preparation to obtain a solution containing 0.1-200mg/ml or 1-200mg/ml AST-3424 crude drug.

The invention further discloses an injection for AST-3424, which is a ready-to-use pharmaceutical composition injection, the composition does not need to be diluted before administration, is suitable for administration in production, and does not need to be diluted or prepared by medical staff in use.

Obviously, the injection for AST-3424 may be a ready-to-use injection in various ways, such as intradermal, subcutaneous, intramuscular, intravenous injection, etc.

The invention discloses an injection for AST-3424 injection, wherein the solvent is water, the solute is composed of AST-3424 bulk drug, an isotonic reagent, ethanol, propylene glycol and a pH regulator, the concentration of the AST-3424 bulk drug of the injection is 0.001-1.000mg/ml, the pH of the injection is 6.8-10.5, and the injection is an isotonic solution.

For adjusting the isotonic agent, any pharmaceutically acceptable osmotic agent can be used with respect to the injectable solution of the present invention. Suitable osmolytes include, but are not limited to, sodium chloride, dextrose, sucrose, fructose, xylitol, glycerol, sorbitol, mannitol, potassium chloride, mannose, calcium chloride, magnesium chloride and other inorganic salts or mixtures thereof in anhydrous or aqueous form.

Isotonic solutions, i.e. 308mmol/L are generally considered to be isotonic for humans, such as: 0.9% physiological saline and 5% glucose solution. Of course, values between 280 and 320mmol/L can also be relaxed and considered as isotonic solutions.

Of course, if the subject to which it is adapted is an animal, such as another primate, the concentration of the isotonic solution is correspondingly adjusted.

Further, the concentration of AST-3424 raw material medicine of the injection is 0.004-0.94mg/ml.

Preferably, the AST-3424 intravenous injection has a pH of 7.4-10.5. The intravenous injection solution at the pH value has good stability, and can ensure that the intravenous injection solution can be stored for a certain time (for example, patients can not take medicine on time temporarily after being prepared on site).

The invention provides an injection for AST-3424, wherein the pH regulator is one or a mixture of sodium citrate, sodium acetate, potassium acetate, monopotassium phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate and potassium carbonate.

An injection for intravenous injection of AST-3424, whose solvent is water, and solute is composed of AST-3424 raw material medicine, glucose, alcohol, propylene glycol and pH regulator sodium bicarbonate, the concentration of AST-3424 raw material medicine of said injection is 0.004-0.94mg/ml, pH of said injection is 7.4, the content of glucose in said injection is 4.5-5.0% by mass, and it is isotonic solution.

The method for preparing the stable AST-3424 injection into the intravenous injection comprises the steps of adding a proper amount of sodium bicarbonate solution of a pH regulator into 5% glucose injection, so that the pH value of the mixed solution is 7.4; adding the stable AST-3424 injection into the mixed solution, and mixing and dissolving.

In the dilution preparation process, the amount and concentration of sodium bicarbonate solution, the amount of 5% glucose injection and the amount of AST-3424 injection of the pH regulator are calculated.

Drawings

Fig. 1 is a flow chart of the process for producing an AST-3424 pharmaceutical formulation.

Detailed Description

One embodiment of the present invention provides a non-ready-to-use concentrated injection.

The disclosed storage stable concentrated form of AST-3424 injection is a concentrated injection of a pharmaceutical composition which needs to be diluted before administration and which needs to be diluted or prepared by medical staff or pharmacist or pharmacy or factory at the time of use as required.

The concentrated injection of the present invention may be diluted with any pharmaceutically acceptable solution of isotonic adjusting agent (as a solute) and water for injection (as a solvent).

The stable AST-3424 injection is a solution containing 1-200mg/ml of AST-3424 bulk drug, and the solvent of the solution is a mixed solvent of monohydric alcohol of C2-C8 and dihydric alcohol or trihydric alcohol of C2-C8.

the O in the injection is greatly reduced after the injection is vacuumized by the protective gas ₂ 、CO ₂ The content of the gas with certain activity is equal, and then inert gas which does not react with the raw material medicine, monohydric alcohol, dihydric alcohol or trihydric alcohol, such as N2 or other inert gases are filled in, and the protective gasWill dissolve in the injection;

The pH adjusting substance includes alkali metal and alkaline earth metal salts of weak acids such as carbonic acid, phosphoric acid, citric acid and acetic acid, and inorganic bases such as alkali metal and alkaline earth metal hydroxides such as Na and K. Some embodiments of the present disclosure demonstrate that certain organic amine organic bases may cause instability of the injection, and although the mechanism of instability is not yet clear, such test results instruct or inspire that it is not desirable to add organic amine organic bases, such as triethylamine, triethanolamine, and other organic amines, to the results of non-rigorous stability tests.

It is obvious that the mixed solvent of the monohydric alcohol of C2-C8 and the dihydric alcohol or the trihydric alcohol of C2-C8 defined in the invention means that the monohydric alcohol and the dihydric alcohol or the trihydric alcohol are respectively liquid at normal temperature and pressure or the mixed solvent is liquid after being mixed.

Obviously, it is also possible that the solvent of the solution is a liquid or semi-liquid mixture of a monohydric alcohol containing C2-C8 and a pharmaceutically suitable water-soluble high molecular polymer. The following description will be made by taking absolute ethanol and absolute propylene glycol as examples.

The 1.0ml injection containing 10mgAST-3424 is a specific example, which is poured into a medium borosilicate glass tube injection bottle (brown light-proof) with a volume of 2ml (or 5 ml), then inert protective gas is poured, a rubber plug is covered and an aluminum plastic combined cover for an antibiotic bottle is sealed.

Excipients used in the present invention include solubilizers, antioxidants (e.g., vitamin E, vitamin C or glutathione), buffers, salts, glucose, stabilizers, electrolytes.

Excipients useful in the present invention include another therapeutic agent, an alkalizing agent, an antibacterial agent, an antifungal agent, and combinations thereof.

The invention meets the requirement of storing stably mixed AST-3424 injection, and the mixed injection is a commercial concentrated product.

In some embodiments, the concentrated injection provided by the invention still has the content of more than 90% after being subjected to accelerated test storage for 6 months at normal temperature (25+/-2 ℃ light-shielding brown penicillin bottle and humidity 60+/-5% RH).

In some embodiments, the concentrated injection provided by the invention still has a content of more than 95% after being subjected to accelerated test storage for 6 months at normal temperature (5+/-2 ℃ light-shielding brown penicillin bottle and humidity 60+/-5% RH).

In some embodiments, the concentrated injection provided by the invention still has the content of more than 98% after being subjected to accelerated test storage for 6 months at normal temperature (-20+/-2 ℃ light-shielding brown penicillin bottle and humidity 60+/-5% RH).

Further, since the AST-3424 drug substance is S-configuration, stability is not only stability of the compound, but also whether chiral inversion occurs during storage (investigating EE value of enantiomeric excess), in investigating the obtained concentrated injection which is relatively stable:

in some embodiments, the concentrated injection provided by the invention still has an EE value of more than 97% after being subjected to accelerated test storage for 6 months at normal temperature (25+/-2 ℃ light-shielding brown penicillin bottle and humidity 60+/-5% RH).

In some embodiments, the concentrated injection provided by the invention still has an EE value of more than 97% after being subjected to accelerated test storage for 6 months at normal temperature (5+/-2 ℃ light-shielding brown penicillin bottle and humidity 60+/-5% RH).

In some embodiments, the concentrated injection provided by the invention still has an EE value of more than 97% after being subjected to accelerated test storage for 6 months at normal temperature (-20+/-2 ℃ light-shielding brown penicillin bottle and humidity 60+/-5% RH).

In particular, in certain embodiments, the stability of the observed injection is inferior to that of an injection without the addition of organic amine (triethanolamine) due to the addition of organic amine (triethanolamine) to the concentrated injection: lower content, higher impurity content and new impurities.

The pH of the compositions of the present invention may be adjusted with an appropriate amount of a pH adjustor comprising an acid-base group. Suitable pH adjusting agents generally comprise at least one acid or salt thereof, or at least one base or salt thereof. Acid-base adjustment is added to achieve the desired pH. For example, if the pH is below the desired pH, a base (or some salt) is added to raise the pH to the desired pH.

Amino acids that can be used in the present invention include, for example, arginine, glycine, methionine, or lysine. In certain embodiments, the amino acid has at least one basic group having a pKa greater than 5, 6, 7, 8, 8.5, or a mixture thereof, or a salt thereof, or a mixture of an amino acid and a salt thereof. The dosage of the amino acid in the invention is 0.1-100 mg/mL, 1-50 mg/mL or 5-25 mg/mL.

In certain embodiments, the concentrated injection solutions of the present invention are sterile, such as by terminal sterilization.

The injection of the composition of the invention is packaged in a pharmaceutically acceptable packaging container, which may be an iv bag or bottle. The infusion bag and the bottle can be made of glass, applicable plastics or polymer materials. The whole or a large part of the area of the packaging container may comprise the following materials: polyvinyl chloride, polyolefin, polyester, polypropylene, or combinations thereof. In other embodiments, only the surface materials that come into contact with the drug injection contain these materials. The invention is preferably a penicillin bottle.

The injection solution provided herein has a sheath (e.g., foil or paper) on the outside to protect the active substance from light. In other embodiments, nitrogen is filled between the injection package and the sheath to prevent oxidation of the composition. In other embodiments, the packaging container (e.g., glass or plastic) may be light resistant.

The concentrated injection solutions of the present invention may also contain an amount of opioid analgesic.

Opioid analgesics may be selected from: alfentanil, allyl rolidine, alfarodin, amprenin, apomorphine, apocodeine, benzomorphan, benzonitrile midete, ibuprofen, buprenorphine, buprenorphine, carfentanil, molamil, codeine, cyclopolychlorinated biphenyl, cipinorphine, dezocine, deanpropylamine, dihydrocodeine, dimesfoado, dimesptanol, dimeshizocine, dioxophenylbutyric acid, dipipperazone, etazocine, ethylthiotin, ethylmorphine, etonizine, fentanyl, hydrocodone, oxymorphone, hydromorphone oxydol, ketomidone, isoprofloxadone, levorphanol, zuo Fenfei alkane, luo Fentai, dolantine, meptazinol, metazocine, methadone, methylmorpholine, metoprolol, mifentinib, morphine-6-glucuronic acid the pharmaceutical composition comprises, as active ingredients, a combination of at least one of meflorphine, nalbuphine, papaverine, nikko, norlevorphanol, normethadone, allylmorphine, orphan/orphan FQ (N/OFQ), normorphine, nopiperidone, oxymethenanthrone, opium, oxycodone, oxymorphone, opium, pentazocine, timolone, fenoxan, phenazocine, phencyclidine, oxymorphone, degranodine, nitenpyramide, pra Luo Gengqin, meperidine, prazidine, propidium, propoxyphene, remifentanil, sufentanil, tapentadol, tramadol, trafentanil, tonidine, nalbuphine; any opioid which is an opioid of phenanthrene, morphinan, phenylmorphine, methadone, phenylpiperidine, propionamidobenzene-4-anilinopiperidine, 4-arylpiperidine, 4-heteroarylpiperidine, or an opioid receptor agonist; any pentacyclic opioid having agonistic activity at the opioid receptor with the same five-nucleus as nalmefene, naltrexone, buprenorphine, levorphanol, meptazinol, pentazocine and dezocine; any fentanyl analog, precursor, derivative or pharmaceutically acceptable salt thereof active at the opioid receptor, and mixtures thereof in racemic or enantiomeric form.

The following examples are presented to aid in the understanding of the invention but are not to be construed as limiting the invention. It is intended that the invention not be limited to the particular embodiments disclosed as the best mode contemplated for carrying out this invention, but that the invention will include all modifications apparent to those skilled in the art upon consideration of the above teachings.

The following are specific experimental and example parts of the invention.

The following experiments will reveal some physicochemical properties of part of the AST-3424 developed by the applicant, which applicant herein states that the following experimental data claims belong to the applicant, in relation to the stability of the concentrated injection solution, i.e. the ready-to-use injection solution, of the present invention.

1. AST-3424 solubility and solution stability study

1.1 buffer/solution preparation

As long as the target concentration is unchanged, different volumes of stock solution and buffer than specified may be used.

And sodium hydroxide solution, 0.2mol/L, 8.00g of sodium hydroxide [ NaOH ] is taken, water is added for dissolution, and water is added for dilution to 1000ml.

Potassium dihydrogen phosphate, 0.2mol/L, potassium dihydrogen phosphate [ KH ] is taken ₂ PO ₄ ]27.22g, dissolved in water and diluted to 1000ml with water.

114.4ml of acetic acid is measured, water is added to dilute the mixture to 1000ml, and the mixture is uniformly mixed.

Boric acid and potassium chloride, 0.2mol/L boric acid [ H ] ₃ BO ₃ ]12.37g and 14.91g of potassium chloride (KCl), dissolved in water and diluted to 1000ml with water.

Acetate buffer at pH 4.5. Taking sodium acetate NaC ₂ H ₃ O ₂ ·3H ₂ 2.99g of O, put into a 1000ml measuring flask, 14.0ml of acetic acid solution is added, then water is added to the scale, and mixed.

phosphate buffer at pH 6.8. 50ml of potassium dihydrogen phosphate solution was placed in a 200ml measuring flask, 22.4ml of sodium hydroxide solution was added, and then water was added to the scale.

phosphate buffer pH 7.4. 50ml of potassium dihydrogen phosphate solution was taken, placed in a 200ml measuring flask, 39.1ml of sodium hydroxide solution was added, and then water was added to the scale.

Alkaline borate buffer at pH 10.0. 50ml of boric acid and potassium chloride solution were taken, placed in a 200ml measuring flask, 43.7ml of sodium hydroxide solution was added, and then water was added to the scale.

1.2 solubility test

An appropriate amount of AST-3424 (hereinafter referred to as drug substance or API) was taken and placed in an appropriate container containing 40ml of the above-described solution (20 ml for organic solvents) until an excess of AST-3424 oil droplets were present in the solution.

The sample was placed in a constant temperature shaking incubator, maintained at 25℃and shaken at a suitable speed (100 rpm). Samples were taken at each predetermined time point, checked for pH, and then centrifuged (10000 rpm,10 minutes) and the corresponding solution or organic solvent (dissolution API for solubility studies) was added to dilute to the appropriate concentration for HPLC analysis (note: dilution ratio after accurate recording test) in accordance with the specifications in Table 1 to obtain solubility data.

If a significantly reduced solubility is observed, it is not necessary to check the solubility for 48h and 72 h.

Table 1: sampling time and solubility test item

Solution/buffer

1h

4h

8h

24h

48h

72h

Ethanol

A

Propylene glycol

A

Sodium acetate buffer at pH4.5

A，P

phosphate buffer at pH6.8

A，P

phosphate buffer at pH 7.4

A，P

Alkaline borate buffer at pH 10.0

A，P

A.P

A，P

Purified water

A，P

Remarks: a=content measurement, measuring the content of AST-3424 in the solution; p=ph

1.3 solution stability study

About 107.32mg of AST-3424 (50% v/v, ethanol) was placed in 50ml volumetric flasks and each was dissolved (organic solvent, buffer solution or purified water) to a scale. At each predetermined time point, 1ml of sample was taken for HPLC analysis. If the API is stable in solutions of different pH, the sampling time point is prolonged, for example, 5 days or longer. Specific sampling times and test items are shown in Table 2 below.

Table 2: sample analysis time for solution stability studies

Solution/buffer

0h

1h

4h

8h

24h

48h

72h

Ethanol

A

Propylene glycol

A

Sodium acetate buffer at pH4.5

A，P

A

A，P

phosphate buffer at pH6.8

A，P

A

A，P

phosphate buffer at pH 7.4

A，P

A

A，P

Alkaline borate buffer at pH 10.0

A.P

A

AvP

Purified water

A，P

A

A，P

Remarks: a=content determination, determining the content of AST-3424 in the solution, HPLC peak purity and total impurities; p=ph

1.4 test methods

For the assay samples of the solubility study, about 1ml of medium was measured, and then centrifuged at 10000rpm for 10 minutes, and the lower clear solution was collected for HPLC analysis. For solution stability studies, samples can be directly injected into HPLC for analysis.

Content was determined using HPLC method: quantification was performed using AST-3424 as an external standard.

UVADD detector wavelength 230nm, C18 column, column temperature 25 ℃.

Mobile phase:

a: 10mmol/L ammonium acetate solution of ammonium acetate in a mixed solvent of 95% water and 5% acetonitrile in volume ratio;

b: 8mmol/L ammonium acetate solution of ammonium acetate in a mixed solvent of 95% acetonitrile and 5% water in volume ratio;

gradient elution was performed.

1.5 test results

The data for solution stability of AST-3424 are summarized in Table 3. Solution stability results of AST-3424 indicated that API was stable for at least 72h in ethanol, ethanol/propylene glycol=1/1, ph7.4 and ph10.0 buffer solutions and at least 24h in ph6.8 solutions at room temperature. The API is unstable in ph4.5 solutions and water, particularly in ph4.5 acetate buffers.

Table 3: solution stability study results of AST-3424 in different solutions

According to the solution stability results, AST-3424 was very unstable in acetate buffer at pH 4.5. Thus, a pH6.8 phosphate buffer, a pH7.4 phosphate buffer, a pH10.0 alkaline borate buffer and purified water were selected for the solubility test. An appropriate amount of AST-3424 to 40mL of medium (8 mL for ethanol/propylene glycol, 50:50V/V) was weighed until an excess of floc was present in the solution. If the solubility of the API in the aqueous solution is > 2% (20 mg/ml), no further API needs to be added.

Table 4: solubility study results of AST-3424

AST-3424 has a solubility in ethanol/propylene glycol (50:50, v/v) solution of greater than 270mg/mL. The solubility of the API was about 23mg/mL in water at pH6.8, pH7.4 and pH10.0, respectively, and about 20mg/mL. The solubility gradually decreases with time due to the instability of the API in ph6.8 and water. At different times, the pH of the solution of the API in the pH6.8, pH7.4, pH10.0 medium remained unchanged. The pH of the aqueous solution containing the API gradually increased from pH5.004 to pH6.512 over 48 hours.

1.6 summary of physicochemical Properties related to injection

Solubility of the polymer. Table 5 summarizes the solubility of AST-3424 in different solvents at 25 ℃. AST-3424 is easily dissolved in alcohol solvents such as ethanol/propylene glycol, and special researchers also primarily examine other monohydric alcohols such as methanol, propanol, butanol, etc. and ethylene glycol, propylene glycol, glycerol, 1, 3-butanediol, 1, 2-butanediol, etc., and the solvents have better solubility for raw materials.

AST-3424 is slightly soluble in water, phosphate buffer pH6.8, phosphate buffer pH7.4 and alkaline borate buffer pH 10.0.

In addition, AST-3424 is stable in phosphate buffer at pH7.4, but is poor in stability in water and phosphate buffer at pH6.8, so the water content should be minimized in the production and storage of the injection (the inventors speculate that the N-containing three-membered ring structure in AST-3424 is likely to undergo ring opening hydrolysis deterioration in the presence of water).

Table 5: AST-3424 solubility

Optical rotation

Optical rotation of AST-3424 [ a ]] ²⁴ _D ＝-30.04°(c＝0.006，EtOH)。

2. Prescription design, preparation and stability study of AST-3424 injection

The formulation, formulation and stability studies were performed by the above solubility and solution stability studies reveal the stability properties of AST-3424 under alkaline conditions.

2.1 prescription design and formulation

AST-3424 pharmaceutical formulation prescription investigation involves the selection of various solvents to prepare different prescriptions.

From the viewpoints of toxicity of the substance, safety of the solvent of the injection, availability, and the like, the inventors selected a prescription solvent composed of ethanol, propylene glycol, triethanolamine (organic amine used for making the pH of the solution alkaline) and the like, and prepared different prescriptions.

Dosages of 10,200 mg/mL were then selected based on solubility to prepare different prescriptions for investigation.

Stability studies of these different formulations were then performed at-20 ℃, 2-8 ℃ and 25 ℃.

HPLC methods were used to monitor the chemical properties of the product, including content, related substances and ee value (enantiomeric excess) to determine the optimal prescription of AST-3424.

Table 6 below shows the different prescription compositions of the 10mg/mLAST-3424 injection

Table 6: different prescription compositions of AST-3424 injection (10 mg/mL)

Table 7 below sets forth the different prescription compositions of 200mg/mLAST-3424 injection

Table 7: different prescription compositions of AST-3424 injection (200 mg/mL)

AST-3424 (dissolved in ethanol) was accurately weighed, placed in a suitable measuring flask, and then different solvents were added to make up the final concentration of the API at 10mg/mL or 200 mg/mL. The different prescription compositions are listed in tables 3 and 4. One milliliter of the bulk solution was measured and poured into a 6ml brown penicillin bottle. After sealing with rubber stopper and aluminum cap, the pharmaceutical preparation is stored for different time periods at-20+ -2deg.C, 5+ -2deg.C or 25+ -2deg.C, dark brown penicillin bottle and humidity 60+ -5% RH. At a predetermined time, the sample is removed and inspected. The test method is the same as the solubility and solution stability of the first section.

2.2 stability test results

At each predetermined time, samples were taken, and HPLC detection was performed on the content of the drug substance in the injection, the relevant substances (i.e., impurities), to obtain the sample schedule analysis contents of tables 8 to 10 and the stability test data of the relevant substances.

Table 8: summary of data for stability at 25℃for different prescriptions (25.+ -. 2 ℃)

Table 9: summary of data for stability at 5℃for different prescriptions (2-8 ℃)

Table 10: summary of data for stability of different prescriptions at-20 ℃ (-22- -18 ℃ C.)

The EE values of the 7 prescriptions, which are relatively stable, were determined and recorded as follows in table 11:

table 11: EE% value for a relatively stable prescription

Remarks: /indicate no test; TBD indicates undetected, ×indicates that the data is bad; ND represents below the instrument detection limit and undetected; N/A means not containing

2.3 results and discussion

Stability results for different formulations of AST-3424 injection are shown in tables 8-11. Stability results for different formulations stored at 25 ℃ it was found that the stability of the formulation increased with increasing proportion of ethanol in the formulation. For formulations containing triethanolamine, the stability of these formulations is lower than formulations without triethanolamine.

F9 is the most stable of the other prescriptions at a dosage specification of 10 mg/ml.

For formulations stored at 2-8deg.C and-20deg.C, the stability of the sample was significantly increased. Samples stored at-20℃are more stable than the formulations stored at 2-8 ℃. The stable prescriptions (i.e., F1 (10 mg/ml), F7 (10 mg/ml), F9 (10 mg/ml), F1-1 (10 mg/ml), F1 (200 mg/ml), F3 (200 mg/ml), and F2-1 (200 mg/ml)) were tested for Enantiomeric Excess (EE), and the EE values for the relatively stable prescriptions are shown in Table 11. The EE values of all the formulations did not change under different storage conditions for 6 months, indicating that no isomer conversion of the active ingredients in these formulations occurred.

By comparing the stability test results of different prescriptions, it can be seen that:

(1) According to the screening research results of the prescription in the table, the stability of the pharmaceutical preparation is improved along with the increase of the proportion of ethanol in the prescription. On the other hand, if triethanolamine is added to the formulation, the pharmaceutical formulation becomes unstable.

(2) The results of the stability study show that the pharmaceutical formulations are more stable in storage at-20 ℃ than at 2-8 ℃ or 25 ℃. The storage temperature has a significant effect on the stability of the pharmaceutical formulation.

(3) The candidate prescription with code F9 was determined to be the most stable of the candidate prescriptions by AST-3424 drug formulation concentration screening. The six month stability results showed no significant change in the relevant material and ee value test items.

(4) Based on the prescription investigation results, prescription F9 (75.0% ethanol and 25.0% propylene glycol) was selected as the final candidate prescription for the AST-3424 pharmaceutical formulation.

3. Light stability study of AST-3424 injection

3.1 sample preparation and detection results

After AST-3424 injection (1 mL:10mg, prescription F9) was prepared, it was placed in 1.1mL clear vials and brown vials, respectively, and sealed. The illumination intensity of the stability test box is measured before the light stability study is started, so that the total illumination intensity of the test sample is ensured to be more than 1.2 multiplied by 10 ⁶ Lux hrs, and the end time of the light stability experiment was calculated (light intensity in the light stability chamber where the sample was located was 5200Lux and required a test period of not less than 10 days). All samples were placed in a light stabilization chamber and sampled at 5, 10 and 20 days to examine the properties, content and related substances of each sample.

The transparent vial is a middle borosilicate glass tube penicillin bottle (pharmaceutical grade), and the brown vial is a middle borosilicate glass tube penicillin bottle (pharmaceutical grade). In the test, the components of the medium borosilicate glass tube and the pH are suitable for the stability under the alkalescence condition of the AST-3424 injection of the invention in consideration of the influence of acid-base property.

The initially prepared AST-3424 injection was placed in a brown vial as a test sample control and stored at-20 ℃ and tested at each sampling time point with the sample stored in the light stabilization chamber. Transparent vials wrapped with aluminum foil were used as a control group for the photostability study and were subjected to sampling test (HPLC method section 1.4) according to the same sampling schedule, and the results are shown in table 12 below.

Table 12: light stability research result of AST-3424 injection

Remarks: content, i.e., the relative percentage content of the corresponding substance tested by HPLC; the related substances, namely impurities in the crude drug API.

3.2 conclusion

The light stability results of the AST-3424 injection stored in the transparent vial indicate that the property of the AST-3424 injection changes from pale yellow to brown when illuminated for 5 days. The active ingredient content of each sampling point is reduced, and the active ingredient content is reduced from 99.9% to 56.6% when the illumination is finished for 20 days.

The stored AST-3424 injecta samples were covered with a clear vial and with aluminum foil and illuminated for 20 days with essentially unchanged active ingredient content and impurities.

AST-3424 injection stored in brown vial, the sample content and related substances remained unchanged when illuminated for 10 days, but the active ingredient content was reduced from 99.9% to 97.3% when illuminated for 20 days.

Light stability studies indicate that AST-3424 injections are unstable under light conditions and should be stored in amber (brown) vials to avoid direct irradiation by the light source.

4. AST-3424 injection temperature cycle stability study

In order to evaluate the stability of the active substance AST-3424 in the temperature cycle, the temperature cycle stability of the AST-3424 injection is researched, and guidance is provided for the storage mode of the product in the transportation, storage and use processes.

Since the second part above has clarified that the optimum storage temperature of the AST-3424 injection is-20 ℃, and when in use, the stored injection needs to be taken out from the low-temperature environment to the normal-temperature 25 ℃ environment, or the injection needs to be put back into the low-temperature environment with the optimum storage temperature of-20 ℃ from the normal-temperature 25 ℃ environment, so that the stability of the injection may be affected by the temperature cycle impact, and therefore, the research on the stability of the injection needs to be carried out.

4.1 test procedure

The prepared AST-3424 injection (1 ml:10 mg) was stored at-20℃for 2 days, and then the sample was transferred to an environment of 2-8℃for two days, and this was taken as a temperature cycle. Three temperature cycles will be performed and the samples will be analyzed after each temperature cycle. The initially prepared AST-2424 injection was stored in a-20℃ refrigerator as a control. The detection items comprise properties, content and related substances. The above process was repeated to perform a temperature cycle from-20℃to 25 ℃.

4.2 test results

Two samples (4 vials) were prepared for each temperature cycle, one sample (2 sample vials) was tested and the other sample (2 sample vials) was ready for use. The cycle times in the original test records are set forth in Table 13, and the results of the temperature cycling stability are summarized in Table 14.

Table 13: temperature cycling study sampling schedule

Vials: indicating the number of drug vials placed in the vial; pull: representing the number of samples

Table 14: temperature cycling stability results of AST-3424 injection

Contraol: a control group; first cycle, second cycle, third cycle: first, second and third cycles

4.3 test results

Three temperature cycling results of the AST-3424 injection from-20 ℃ to 2-8 ℃ show that the content and impurities of the sample are not changed during the period. The results of the temperature cycle test of the AST-3424 injection from-20 ℃ to 25 ℃ show that the content of the related substances in the sample is not changed, and the content of the related substances is only slightly increased (1.3% is changed to 1.4%). The above results indicate that AST-3424 injection is stable over three temperature cycles of-20℃to 2-8℃and-20℃to 25 ℃.

5. Preparation process and specific examples of concentrated injection

The process flow diagram of the AST-3424 pharmaceutical preparation is shown in figure 1.

Step 1: dissolving and mixing

Step 2-1: adding ethanol solution

The prescribed amount of AST-3424 drug substance (pyrogen removed) was weighed out in a beaker and put into a formulation tank. Adding 50% of medicinal absolute ethanol (pyrogen removed) and stirring until dissolving (dissolving time 15min, stirring speed 50HZ 50 rpm).

Step 2-2: propylene glycol is added

The prescribed amount of propylene glycol (pyrogen removed) was added and stirred until dissolved (dissolution time 15min, 50HZ, i.e. 50 revolutions per minute).

Step 2-3: mixing

Adding medicinal anhydrous ethanol (pyrogen removed) of 50% of the prescription amount, stirring until dissolving (dissolving time 15min, stirring speed 50HZ 50 rpm).

Step 3: degerming

And (3) performing sterilization operation on the solution obtained in the step (2).

Step 4: aseptic filling

Sterile filling is carried out with a filling quantity of 1.0-1.2ml (0.860-1.032 g).

Step 5: roll cover and appearance inspection

The filled medicine bottles are conveyed to a cover rolling room through a conveying net belt for cover rolling. Appearance inspection is performed.

Step 6: pass inspection

The AST-3424 injection was sampled for QC testing and stored at-20℃for clinical use after QA release.

The formulation design, stability and other properties of the concentrated injection and specific preparation examples were studied above, and the preparation process of the ready-to-use injection and stability study will be disclosed below.

6. Compounding and stability study of ready-to-use intravenous solutions

In order to provide compatibility information of AST-3424 injection and compatible solution, short-term use stability of the injection in a common intravenous diluent of 5% glucose injection is required to be examined. Considering that API is unstable in an acidic environment, the pH of 5% dextrose injection was first adjusted to 7.4 with sodium bicarbonate solution.

6.1 Experimental methods and Processes

Preparation of 5% glucose injection

To a 5% dextrose injection (250 mL:12.5 g) was added 0.22mL sodium bicarbonate solution using a 1mL syringe and mixed well, at which point the pH of the mixed solution was about 7.4.

Preparation of AST-3424 compound solution

AST-3424 injection (1 mL:10mg, clinical batch number: 20170701) and 5% glucose injection were used to prepare the compound solutions at concentrations of 0.004mg/mL, 0.071mg/mL and 0.94 mg/mL.

Preparation of 0.004mg/mL Compound solution

A single AST-3424 injection (1 mL:10 mg) was taken, and 0.1mL was withdrawn with a 1mL syringe and added to a mixing bag containing 250mL of 5% dextrose injection (pH adjusted to 7.4 with pH adjuster). And ensures uniform mixing of the mixed solution by turning the mixing bag over multiple times before storage and sampling.

Preparation of 0.071mg/mL Compound solution

1.8mL of the sample was withdrawn from the two bottles of AST-3424 injectate using a 2.5mL syringe and added to a mixing bag containing 250mL of 5% dextrose injectate (previously pH adjusted to 7.4 with pH adjuster). And ensures uniform mixing of the mixed solution by turning the mixing bag over multiple times before storage and sampling.

Preparation of 0.94mg/mL Compound solution

A26 mL sample was withdrawn from a 26 bottle AST-3424 injection using a 30mL syringe and added to a mixing bag containing 250mL of 5% dextrose injection (previously pH adjusted to 7.4 with a pH adjustor). And ensures uniform mixing of the mixed solution by turning the mixing bag over multiple times before storage and sampling.

Temperature and illumination conditions

All the compound solution samples were placed under natural light conditions and stored at room temperature (25.+ -. 3 ℃) for 24 hours.

Sampling time point and test item

Two 10mL samples of the reconstituted solution were taken in each bag using 20mL syringes at 0, 1, 2, 4, 8 and 24 hours, respectively, one for testing and the other as a backup. The test items are as follows:

1) Visual inspection;

2) Osmotic pressure (initial and final sampling time points);

3) Compounding the pH value of the solution;

4) Content (tested using test method in section 1.4);

5) Related substances (tested using the test method in section 1.4);

6)e.e.

Table 15: AST-3424 compound solution sampling time point and test item

Remarks I = visual inspection; o = osmotic pressure; p=ph; a = content; r=related substance; e=e.e. value.

6.2 experimental results

All experimental data will be recorded in tables 16-20 summarizing stability data for AST-3424 formulated solutions diluted to different concentrations.

Table 16: stability data of compound solution of injection (1 mL:10 mg) with concentration of 0.004mg/mLAST-3424 and 5% glucose injection are summarized

Time: 07MAR2018-08MAR2018 temperature: relative humidity at 25.6-26.2 ℃:26.8-33.8%

Table 17: stability data of compound solution of injection (1 mL:10 mg) with concentration of 0.071mg/mLAST-3424 and 5% glucose injection are summarized

Time: 08MAR2018-09MAR2018 temperature: 23.9-26.6 ℃ relative humidity: 18.1-28.9%

Table 18: data on substances of compound solution of injection (1 mL:10 mg) with concentration of 0.071mg/mLAST-3424 and 5% glucose injection are summarized

Table 19: stability data of compound solution of injection (1 mL:10 mg) with concentration of 0.94mg/mLAST-3424 and 5% glucose injection are summarized

Time: 09MAR2018-10MAR2018 temperature: 22.1-27.2 ℃ relative humidity: 18.1-25.2%

Table 20: data on substances of injection (1 mL:10 mg) and 5% glucose injection compound solution with concentration of 0.94mg/mLAST-3424 are summarized

Remarks:

regarding the content of the compounding solution

1) The preparation of the 0.004mg/mL compound solution requires that the volume of AST-3424 injection is 0.1mL, but 0.1mL sample is not easy to be accurately measured by a 1mL syringe. 0.0055mg/mL is the actual measurement at the initial time point, and its value is about 138% of the theoretical value;

2) The actual volume of the 5% glucose injection was about 268mL and the theoretical volume was 250mL, so that the actual results at the initial time points were 0.067mg/mL and 0.86mg/mL, respectively, when samples were prepared at concentrations of 0.071g/mL and 0.94 g/mL.

3) PRT, retention time in minutes, the retention time of the chromatographic peak of a corresponding impurity was detected in the HPLC method section 1.4, which corresponds to a certain impurity.

4) Initial, initial value.

6.3 conclusion of the test

1) The compound solution with the concentration of 0.004mg/mL and 0.071mg/mL is colorless transparent solution, and the compound solution with the concentration of 0.96mg/mL is yellowish transparent solution.

2) The osmotic pressure of the low concentration (0.004 mg/mL) compound solution is lower than the isotonic pressure, and the compound solution is almost isotonic when the concentration is 0.071mg/mL (284-288 mmol/kg) (0.074 mg/mL is isotonic solution). The high concentration (0.94 mg/mL) of the compound solution was hypertonic (561-571 mmol/kg). 0. The osmolarity data for the 8 and 24h sampling points indicated that the osmolarity of the reconstituted solution remained constant for 24h at the three concentrations.

3) During the 24h study, the pH of the compounded solutions at concentrations of 0.004mg/mL, 0.071mg/mL and 0.94mg/mL fluctuated very little during storage.

4) The content measurement results show that the compound solutions with the concentrations of 0.004mg/mL, 0.071mg/mL and 0.94mg/mL are stable in the storage period of 24 hours.

5) Regarding the substances, the total impurities of the compound solutions with the concentrations of 0.071mg/mL and 0.94mg/mL are not more than 2.0% in 24 hours, and remain substantially unchanged at room temperature (25.+ -. 3 ℃) for 8 hours. However, the total impurities in the compound solution with the concentration of 0.94mg/mL are increased within 24 hours. A new impurity peak was observed at concentrations of 0.071mg/mL and 0.94mg/mL after 8 hours and 4 hours of storage, respectively, at a level of about 0.26-0.35%. In addition, a new impurity peak was observed at a concentration of about 0.10-0.13% in the 24h storage of the 0.071mg/mL and 0.94mg/mL of the compounded solution. The compound solution at low concentration (0.004 mg/mL) was not assayed for the relevant substances due to the AST-3434 content being below the limit of quantification.

6) At both concentrations of 0.071mg/mL and 0.94mg/mL, the e.e. value of the compounded solution remained unchanged over a 24h storage period. The low concentration of the compounded solution (0.004 mg/mL) did not meet the requirements of the method, and no e.e. value was measured.

7) The results show that under the conditions of natural light and room temperature (25+/-3 ℃), the compound solution (concentration range) of AST-3424 injection (1 mL:10mg preparation) and 5% glucose injection (with pH value adjusted to 7.4: 0.004mg/mL-0.94 mg/mL) is preferably used within 8 hours.

In the case of compounding, according to the experimental conclusion that the AST-3424 solution is stable under the alkaline condition of pH in the above experiment, it is necessary to adjust 5% glucose injection (pH 3.2-6.5 specified in pharmacopoeia) or physiological saline (pH 4.5-7.0 specified in pharmacopoeia) to pH6.8-10.5, preferably slightly alkaline, preferably 7.4, by using alkaline solution such as NaHCO3, so that the injection can be prepared in a qualified and field-used preparation process after the concentrated AST-3424 injection preparation is added subsequently.

Claims

1. A stable AST-3424 injection preparation is a solution containing 0.1-200mg/ml of AST-3424 bulk drug, the solvent of the solution is a mixture of ethanol and propylene glycol, wherein the volume ratio of the ethanol to the propylene glycol is (70% -75%): (30% -25%), and the volume ratio does not include 70%:30% and 75%:25%.

2. The stable AST-3424 injectable formulation of claim 1, which is a solution containing 1-200mg/ml of AST-3424 drug substance.

3. The stable AST-3424 injectable formulation of claim 2, which is a solution containing 10mg/ml of AST-3424 drug substance.

4. The stable AST-3424 injection formulation of claim 1, which is free of added water, the water content being controlled within 0.5% by mass.

5. The stable AST-3424 injection preparation product comprises a packaging container and an AST-3424 injection preparation contained in the container, wherein the injection preparation is an injection preparation as set forth in any one of the above claims 1-4, wherein the AST-3424 bulk drug content in the product is 1-200mg, and the injection preparation is filled in a light-resistant glass bottle made of medium borosilicate glass filled with protective gas.

6. The product of claim 5, wherein the AST-3424 drug substance is present in the product at a level of 10mg.

7. The product of claim 5, wherein the AST-3424 drug substance is present in the product at 20mg.

8. A process for the preparation of a stable AST-3424 injectable formulation according to any one of claims 1 to 4, comprising the following operations: dissolving AST-3424 raw material medicine and ethanol for the first time; adding propylene glycol with the prescribed amount for the second dissolution preparation to obtain a solution containing 0.1-200mg/ml AST-3424 bulk drug.

9. The method of claim 8, wherein a solution of 1-200mg/ml AST-3424 drug substance is obtained.

10. A process for the preparation of a stable AST-3424 injectable formulation according to any one of claims 1 to 4, comprising the following operations: dissolving AST-3424 raw material medicine and partial ethanol with a prescription amount for the first time; adding the propylene glycol with the prescription amount for dissolving and preparing for the second time; adding the rest ethanol with the prescription amount, mixing and dissolving to obtain solution containing 1-200mg/ml AST-3424 crude drug.

11. An injection for AST-3424 injection, the solvent of which is water, the solute comprises an AST-3424 bulk drug, an isotonic reagent, ethanol, propylene glycol and a pH regulator, the concentration of the AST-3424 bulk drug of the injection is 0.001-1.000mg/ml, the pH of the injection is 7.4, and the injection is an isotonic solution, wherein the volume ratio of the ethanol to the propylene glycol is (70% -75%): (30% -25%), and the volume ratio does not include 70%:30% and 75%:25%.

12. The injection for AST-3424 of claim 11, wherein,

the concentration of AST-3424 raw material medicine of the injection is 0.004-0.94mg/ml;

the pH regulator is one or more of sodium citrate, potassium citrate, sodium acetate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate and potassium carbonate;

The isotonic agent is sodium chloride, glucose, sucrose, fructose, xylitol, glycerol, sorbitol, mannitol, potassium chloride, mannose, calcium chloride or magnesium chloride.

13. An injection for AST-3424 intravenous injection, the solvent of which is water, the solute is composed of AST-3424 raw material medicine, glucose, ethanol, propylene glycol and pH regulator sodium bicarbonate, the concentration of the AST-3424 raw material medicine of the injection is 0.004-0.94mg/ml, the pH of the injection is 7.4, the content of glucose in the injection is 4.5-5.0% by mass, and the injection is an isotonic solution, wherein the volume ratio of ethanol to propylene glycol is (70% -75%): (30% -25%), and the volume ratio does not include 70%:30% and 75%:25%.

14. The method for preparing the stable AST-3424 injection into the intravenous injection comprises the steps of adding a proper amount of sodium bicarbonate solution of a pH regulator into 5% glucose injection, so that the pH value of the mixed solution is 7.4; adding the stable AST-3424 injection preparation of any one of claims 1-4 or the stable AST-3424 injection preparation product of any one of claims 5-7 into the mixed solution, and mixing and dissolving.